The present disclosure relates to a microscopy system, a microscopy method, and a non-transitory computer readable storage medium storing a microscopy program for observing a subject through images acquired by a microscope device.
For observation of a subject having depth, such as cell nuclei or stem cells, by using a biological microscope having a focal depth of several tens of μm level, users need to promptly identify the site of interest that is present in a depth direction (Z direction) along the optical axis of a microscopy optical system. To meet the users' need, there is a method with which capturing is sequentially conducted while shifting a focal plane of the microscopy optical system along the optical axis to acquire and observe images with different focal planes. Such images acquired with different focal planes are also collectively referred to as Z-stack images.
Capturing Z-stack images is time-consuming because multiple images need to be acquired. Furthermore, for fluorescence observation, when the subject is continuously irradiated with excitation light, “discoloration” which is a reduction in the intensity of fluorescence occurs; therefore, it is preferable to conduct desired capturing in as short period of time as possible and as few times as possible. Furthermore, as the exposure time for fluorescence observation is often longer than that for bright-field observation, the time during which the subject is irradiated with excitation light is inevitably long when Z-stack images are acquired.
Therefore, as the technology for acquiring images with different focal planes, sweep capturing is known which is capturing by moving the subject or the imaging device in the Z direction during exposure (for example, see International Publication WO2012/105222 and Japanese Laid-open Patent Publication No. 2012-013804). On sweep images, a structure that is present in the Z direction is superimposed, and therefore users are allowed to know the presence of a structure with depth on a two-dimensional plane perpendicular to the Z direction on the basis of sweep images. Furthermore, omnifocal images may be acquired by performing restoring processing on sweep images; thus, as compared with a method for acquiring omnifocal images from Z-stack images, a capturing time may be shortened. Furthermore, Japanese Laid-open Patent Publication No. 2012-013804 discloses the configuration in which the sweep capturing is applied to microscopes.